Objective To select the optimal alcohol extraction process for the classic prescription Xuefu Zhuyu Decoction (XZD, 血府逐瘀汤), enabling the retention of small-molecule substances in traditional aqueous extracts, thereby providing experimental basis for the development of transdermal formulations. Methods The contents of 12 active components (adenosine, catalpol, amygdalin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, liquiritin, narirutin, naringin, hesperidin, neohesperidin, glycyrrhizic acid) in XZD, the extract yield, and the fingerprint similarity were used as critical quality attributes (CQAs). Ethanol concentration, extraction time, and solvent ratio were selected as critical process parameters (CPPs), and experiments were conducted using a Box-Behnken design-response surface methodology (BBD-RSM). The benchmark correlation degree of each experimental sample index was calculated, and the weights of each CQA were determined using the entropy weight method to compute a comprehensive score. The TOPSIS model was applied to calculate the relative closeness of each sample to the benchmark sample and rank them, with the optimal alcohol extraction process screened based on the comprehensive score. Additionally, ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) was used to description the chemical components of the benchmark sample and the optimal alcohol extraction process sample, further verifying retention effect of small-molecule components.. Results Analysis using the entropy weight-TOPSIS model determined the optimal alcohol extraction process for XZD as follows: ten times the amount of 60% ethanol, with each extraction lastin 3 h. The average comprehensive score of three validation samples reached 0.501, with RSD of 0.75%, which met the requirements and confirmed that the process was stable and the method was applicable. Meanwhile, UPLC-Q-TOF-MS/MS analysis showed that the main chemical composition of the optimal process sample were similar to those of the benchmark sample. Conclusion The comprehensive screening strategy based on the benchmark correlation degree and entropy weight-TOPSIS model is feasible and effective. The obtained XZD alcohol extraction process parameters are robust and reliable, and the small molecule substances of the benchmark decoction can be effectively retained. The chemical substance basis is similar to that of the benchmark decoction. This study can provide a reference for the research and development of XZD transdermal formulations.

Xuefu Zhuyu Decoction  /  quality by design  /  standard relation  /  entropy weight-TOPSIS model  /  fingerprint  /  Box-Behnken design response surface methodology  /  UPLC-Q-TOF-MS/MS